Compositions and method for the treatment of vascular spasms



United States Patent 3,165,444 CQh/lltlfililfiNS AND METHGD FQR THE TREATMENT 91F VASCULAR diASME Daniel iiertin, Montrouge, France, assignor to Roussel- UCLAF, iaris, France, a corporation of France No Drawing. Filed Apr. 17, 1962, Ser. No. 188,226 Claims priority, application France, Apr. 22, 1%1, Patent 859,596 6 Claims. (Cl. lei-65) The invention relates to a novel method of treating vascular spasms by administration of l'-(5fi-androstane- 3u-ol-ll-one-lLB-yl)-3',5',8-trioxabicyclo- (2,2,2) .octane and its derivatives. The invention also relates to novel compositions for the treatment of vascular spasms.

In commonly assigned, copending patent application,

Serial No. 172,039, filed lanuary 23, 1962, now US. Patent No. 3, )79,385,- l-(5,6-androstane-3a ol ll onel75-yl)-3,5',8-trioxabicyclo-(2,2,2)-octane and its esters are disclosed as intermediates for the preparation of 3aacetoXy-ZO-bis-(hydroxymethyl)-55prcgnane-2l ol 1l-' one. it has now been discovered that the said compound and its 3a-estcrs and others have interesting pharmacological properties. 1

l'-(5B-androstane-3o-ol-ll-one-l7fl-yl)-3',5',8- trioxabicyclo-(2-,2,2)-octane possesses a coronary dilatory activity and a spasmolytic activity particularly marked on vascular spasms. It can be used for treatment of spasms of coronary origin, arterial or venous spasms. The cesters and others possessthe same activity since they are R O e wherein R is selected from the group consisting of hydrogen, an acyl radical of an organic carboxylic acid having 1 to 18 carbon atoms and a lower'alkyl radical and a pharamaceutical carrier.

The acyl radical of the organic carboxylic acid having 1 to 18 carbon atoms may be derived from an aliphatic, aromatic, cycloaliphatic or heterocyclic carboxylic acid. Examples of suitable acids are alkanoic acids such as formic. acid, acetic acid, propionic acid, butyric acid, iso butyric acid, valeric acid, isovaleric acid, trimethyl acetic acid, caproic acid, ,B-trimethyl propionic acid, heptanoic acid, caprylic acid, pelarginic acid, capric acid, undecylic acid, lauric acid, myristic acid, palmitic acid and stearic acid; alkenoic acids such as undecylenic acid and oleic acid; cycloallryl carboxylic acids such as cyclopentyl car boxylic acid, cyclopropyl carboxylic acid, cyclobutyl carboxylic acid and cylohexyl carboxylic acid; cycloalkyl alkanoic acids such as cyclopentyl acetic acid, cyclohexyl acetic acid, cyclopentyl propionic acid and cyclohexyl propionic acid; arylallianoic acids such as phenyl acetic acid and phenyl propionic acid; aryl carboxylic acids such "as benzoic acid benzoic acid; pheuoxy alkanoic acids such as phenoxy the invention to provide a novel $355,444 Patented Jan. 12, 1965 "ice acetic acid, p-chlorophenoxy acetic acid, 2,4dichl0rm phenoxy acetic acid, 4-terbutylphenoxy acetic acid, 3- phenoxy propionic acid and 4-phenoxy butyric' acid; heterocyclic carboxylic acids such as furane-Z-carboxylic acid, 5-tter-butylfurane-Z-carboxylic acid, S-bromofurane- Z-carboxylic acid and nicotinic acids; B-ketoalkanoic acids, such as acctylacetic acid, propionyl-acetic acid and butyrylacetic acid; amino acids such as diethylaminoacetic acid and aspartic acid.

The compositions of the invention may be prepared in the form of injectable solutions or suspensions, prepared in ampules, in the form of multiple-dose flacons, in the form of tablets, in the form of aromatized suspensions and in the form of suppositories according to the usual methods. The useful individual dose is controlled between 10 and 50 mg.

The compounds of Formula I are prepared as described in the said copending application Serial No. 172,039 by esterifying ZO-bis-(hydroxymcthyD-SB pregnane 3a,2ldiol-ll-one with an alkyl orthoformate such as ethyl orthoformate in the presence of an acid catalyst such as p-toluene sulfonic acid to form 1-(5fi-androst ane-3a-ol- 1l-one-17/3-yl)-3',5,8-trioxabicyclo-(2,2,2)-octane which may then be subjected to reaction with an acylating agent such as acetic acid anhydride to form the corresponding 3a-acyloxy compound or with a lower alkylation agent to form the corresponding S-lower, alkoxy compound. The reaction scheme is illustrated in Table I.

* radical.

and 2,4-dinitrog The 20-bis-(hydroxymethyl)-5,6-pregnane 35;,21 diolll-one is prepared according to the process of copending,

commonly assigned application Serial No. 174,692, filed February 21, 1962, now Patent No. 3,120,518,which is described in Comptes Rendus de IAcademie des Sciences, vol. 254 (1962), pp. 42-46. The reaction scheme is summarized in Table II.

7 3 TABLE II 01: arn-H0 V (IJHO A oHo O/=IU=CH ojClUrHz g A00 7% I AcO-- ornoH O--GH;OH

onion CHZOH v CH OH wherein Ac is an acyl radical such as acetyl.

As examples of the compounds used in the composition, there is 1-(5B-androstane-3a-ol-l1-one17;3-yl)-3',5',8'- trioxabicyclo-(2,2,2)-octane which occurs in the form of a colorless compound, soluble in benzene and chloroform, slightly soluble in ether and insoluble in water and which has an instantaneous melting point determined on the Kofler block of 230 C.

AnalysiS.-C H O (molecular weight=404.53): Calculated: C, 71.25%; H, 8.97%. Found: C, 71.5%; H, 9.1.

Infrared spectra (chloroform) is in accord with the structure.

Another example is 1'-(3e-acetoxy-5B-androstane-11- one 17B yl) -3,5,8-trioxabicyclo-(2,2,2)-octane which occurs in the form of a solid colorless compound, soluble in benzene and chloroform, slightly soluble in alcohol and 7 ether, very slightly soluble in aqueous propylene glycol and insoluble in water and which has an instantaneous melting point determined on the Kofler block of 243 C.

The method of the invention comprises administering an elfective amount of a compound of Formula I daily. "The compounds of Formula I may be administered by, oral, perlingual, transcutaneous or rectal methods. The

useful daily dosage'varies from 25 to 100 mg. depending upon the method of administration. i g

In the following examples there are described several i 7 preferred embodiments to illustrate the invention. However, it; should be understood that the invention is not intended to be limited to the specific embodiments.

Pharmacological Study of 1 56-14 ndr0stane-3otOl-11- 0116-1 7 p-Yl -3,5 ,'8'-T rioxabicyclo- (2,2,2 -Octane A. DETERMINATION OF THE SPASMOLY'IIC EFFECT ON THE CONTRACTION OF THE SMALL INTESTINE OF GUINEA PIGS An isolated intestinal loop was suspended in an isolated organ container of 10 cc. containing Tyrode liquid maintained at 37 C. and constantly oxygenated.

(a) Contracture by barium -chl0ride.T he contracture of the intestine was provoked by 200 to 300 gammas per cc. of barium chloride." '1'-(5,6-'androstane-3aol-l l-one thebath at the maximum of the contracture and the minimum concentration provoking partial decontraction and total decontraction was determined. A concentration of 5 gammas per cc. of the product provoked a progressive decontraction of aboutf50%. A concentration of to gammas per cc. brought about a total-decontraction.

new addition of acetylcholine.

(c) Contractni e by histamine-Thecontraction of the intestine was obtained by the addition of 0.005 gamma per cc. of histamine. A total decontraction was obtained by the addition of 5 'gammas per cc, of the compound.

. This same concentration caused the inhibition of the contracting effect of histamine.

I B. DETERMINATION OF THE SPASMOLYTIC EFFECT ON THE ISOLATED BILE VESICULE OF GUINEA PIGS This test was made under the conditions described by Chiray et al., .Revue du Foie, 1943 (2nd series), and by using the vesicule of guinea pigs fasting for a period of 24 hours. The contracture Was provoked by the addition of barium chloride in a concentration of 20 gammas per cc. 1'-(5B-androstane-3a-ol-ll-one-l75-yl)-3',5,8' trioxabicyclo-(2,2,2)-octane inhibited this spasm at a concentration of 1 to 2 gammas per cc. 1

C. ACTION ON CORONARYBLOOD FLOW The study of the action of the compound-on coronary blood flow was made on the isolated rabbit heart by using a technique inspired by Langendorf (Arch. gesam.

Physiol, 1895, 61, 291). In this method, the. heart was suspended by the aorta to a cannula andthe coronary system was perfused by means of this cannula under a tration desired.

On aproper apparatus, the coronary blood how and the ventricular contractions were registered, The threshold concentration of the compound which clearly augmented the blood flow of such a preparation was systematically searched for and the table below furnishes the results obtained with'this compound, as Well as with trinitrine and papaverine under the same experimental con ditions.

V This effect was durable and resisted repeated washings a, 1 e e, 444. 5

TABLE III Threshold Eflects on ventricular active eoncen- Increase in contractions tration in g. coronary blood Duration Compounds studied per co. in the flow in percent of action perfusion of normal in minutes On the On the liquid blood flow amplitude, frequency,

percent percent 1 10 2 to 20 O 5 Papaverine 10 20 15 0 l-(B-androst 1 0. 01 to 0.1 20 15 to 20 7 yl)-3,5,8-trioxabicyclo-(2,2,2)- octane D. DETERMINATION OF THE VASOMOTOR EFFECT ON THE ISOLATED RABBIT PAW This test was made according to a technique described by Binet et al. (Presse Medicale 1949, p. 445). The blood of an animal anesthetized with urethane was drawn from the carotid and re-injected by means of a Jouvelet transfusion apparatus under a constant blood flow into the femoral artery of the isolated paw. A manometer attached to the outlet of the transfusion apparatus permitted the registration of the pressure in the paw which gives a reading corresponding to the degree of the vasodilation or vasoconstriction. The beginning of the vasoconstruction was provoked by the injection of 5 mg./kg. of barium chloride.

1'-(5 8-andr0stane-3a-ol-1l one 17B yl) 3',5',8'-trioXabicyclo-(2,2,2)octane was dissolved in a concentration of 10 mg./cc. in a solvent miscible in Water. The effect of the said compound on the peripheric circulation could be seen at a dose of l mg./kg. injected intravenously. A second injection of the same dose effected an important and long lasting lowering of the arterial pressure in the isolated paw so that the arterial pressure returned almost to the value which it had before the injection of barium chloride.

Acute Toxicity The toxicity tests were made on mice of the Rocldand strain weighing between 18 and 22 grams. 1'-(5,B-an- 'drostane-3a-ol-11-one 1718 yl) 3,5,8' trioxabicyclo- (2,2,2)-octane was utilized in suspension containing 10 mg./ cc. in distilled water containing polysorbate 80. It was injected in this form, after dilution in the case of the slighter dose, by subcutaneous methods into two groups of ten mice at doses of 50 and 100 mg./kg., respectively, in a volume of 0.2 cc. per 20 grams of mice.

The animals were held under observation for a period of 48 hours. No symptoms of intoxication nor mortality were noted in the course of this period. l-(5fi-andro stane-3a-ol-l1-0ne-17fl-y1)-3,5',8'-trioxabicyclo (2,2,2)- octane is thus devoid of toxicity even at a dose of 100 mg./kg. administered to mice by subcutaneous methods.

Various modifications of the compositions and method of the invention may be made without departing from the spirit or scope thereof, and it is to be understood that the invention is to be limited only as defined in the appended claims.

I claim:

1. A method for controlling vascular spasms which comprises administering to an animal having vascular spasms 25 to mg. of a compound having the formula CHZO wherein R is selected from the group consisting of hydrogen, lower alkyl and an iacyl radical of an organic carboxylic acid having 1 to 18 carbon atoms and a pharmaceutical carrier.

5. The composition of claim 4 wherein the compound is 1-(3a-acetQXy-SB-androStane-ILone-17f! yl) 3',5,8'- trioxabicyclo- (2,2,2) octane.

6. The composition of claim 4 wherein the compound is 1-(5[i-and rostane-3a-ol 11 one 17 3 yl) 3,5',8'- trioxabicyclo-(2,2,2)-octane.

References Cited by the Examiner UNITED STATES PATENTS 2/63 Bertin et al. 167-65 12/63 Bertin et a1 167--65 LEWIS GQTTS, Primary Examiner.

FRANK CACCIAPAGLIA, JR., Examiner. 

1. A METHOF FOR CONTROLLING VASCULAR SPASMS WHICH CMOPRISES ADMINISTERING TO AN ANIMAL HAVING VASCULAR SPASMS 25 TO 100 MG. OF A COMPOUND HAVING THE FORMULA 